Benke Dietmar, Möhler Hanns
University of Zurich, Zurich, Switzerland.
Curr Protoc Pharmacol. 2006 Dec;Chapter 1:Unit1.16. doi: 10.1002/0471141755.ph0116s35.
This unit describes ligand binding assays for the characterization of the benzodiazepine (BZ) site of GABA(A) receptors in the central nervous system. The Basic Protocol provides a procedure for analyzing benzodiazepine binding to GABA(A) receptors from brain. The experimental design of the method is suitable for all commercially available radioligands that interact with the BZ site of central GABA(A) receptors. By using the appropriate radioligand in combination with particular displacer compounds, individual (or groups of) GABA(A) receptor subtypes can be identified. Preparation of membrane suspensions for this procedure is described in a Support Protocol, and five Alternate Protocols are provided for the identification of diazepam-sensitive GABA(A) receptors, the α1-subtype of diazepam-sensitive receptors, the α5 subtype of diazepam-sensitive receptors, both diazepam-sensitive and diazepam-insensitive receptors, and diazepam-insensitive receptors exclusively. Each protocol contains a brief description of the requirements for radioligand binding to these five types of BZ sites.
本单元描述了用于表征中枢神经系统中GABA(A)受体苯二氮䓬（BZ）位点的配体结合测定法。基本方案提供了一种分析苯二氮䓬与脑内GABA(A)受体结合的程序。该方法的实验设计适用于所有与中枢GABA(A)受体BZ位点相互作用的市售放射性配体。通过使用适当的放射性配体与特定的置换化合物相结合，可以鉴定出单个（或一组）GABA(A)受体亚型。本程序的膜悬浮液制备方法在支持方案中进行了描述，并提供了五个替代方案，分别用于鉴定地西泮敏感的GABA(A)受体、地西泮敏感受体的α1亚型、地西泮敏感受体的α5亚型、地西泮敏感和不敏感的受体以及仅地西泮不敏感的受体。每个方案都简要描述了放射性配体与这五种类型BZ位点结合的要求。
